LOCUS       sequence-174853-        6082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6082)
  AUTHORS   Nguyen HT, Bride SH, Badawy AB, Adam RM, Lin J, Orsola A, Guthrie 
            PD, Freeman MR, Peters CA
  TITLE     Heparin-binding EGF-like growth factor is up-regulated in the 
            obstructed kidney in a cell- and region-specific manner and acts to 
            inhibit apoptosis.
  JOURNAL   Am J Pathol. 2000 Mar . 156(3):889-98.
  PUBMED    10702406
REFERENCE   2  (bases 1 to 6082)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Am J 
            Pathol. 2000 Mar . 156(3):889-98."
COMMENT     Sequence Label: pcDNA3 proHB-EGF WT
FEATURES             Location/Qualifiers
     source          1..6082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    119..581
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(175..192)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     402..421
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             620..1639
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             620..1639
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     primer_bind     complement(1949..1966)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1955..2179
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2226..2245
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2305..2321)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2305..2321)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2318..2340)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2329..2345
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2353..2359,2360..2377,2378..2383))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2398..2419
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2536..2553)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2707..3295)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2787..2806)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3466..4257,4258..4326))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4089..4108
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4327..4431)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4506..4525)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4697..5076
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5077..5280
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5231..5251
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5325..5344
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5325..5343
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6008..6037
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
ORIGIN
        1 cccccccccc stctccctnc ccccccccta acgttactgg ccgaagccgc ttggaataag
       61 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
      121 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
      181 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
      241 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
      301 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
      361 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
      421 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
      481 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
      541 accacgggga cgtggttttc ctttgaaaaa cacgatgata agcttgccac aacccgtacc
      601 aaagatggat agatccggaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
      661 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
      721 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
      781 tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga
      841 agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca
      901 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
      961 ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt
     1021 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
     1081 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
     1141 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     1201 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     1261 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
     1321 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     1381 tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     1441 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     1501 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt
     1561 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     1621 tccgagggca aaggaataga gtagatgccg accgaacaag agctgatttc gagaacgcct
     1681 cagccagcaa ctcgcgcgag cctagcaagg caaatgcgag agaacggcct tacgcttggt
     1741 ggcacagttc tcgtccacag ttcgctaagc tcgctcggct gggtcgcggg agggccggtc
     1801 gcagtgattc aggcccttct ggattgtgtt ggtccccagg gcacgattgt catgcccacg
     1861 cactcgggtg atctgactga tcccgcagat tggagatcgc cgcccgtgcc tgccgattgg
     1921 gtgcagatct ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     1981 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2041 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     2101 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     2161 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc gagtgcattc
     2221 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     2281 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2341 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     2401 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2461 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2521 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2581 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2641 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2701 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2761 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2821 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2881 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2941 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3001 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3061 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3121 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3181 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaaaaaacca ccgctggtag
     3241 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3301 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3361 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3421 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3481 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3541 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3601 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3661 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3721 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3781 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3841 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3901 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3961 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4021 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4081 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4141 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4201 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4261 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4321 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4381 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4441 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
     4501 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     4561 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     4621 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
     4681 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
     4741 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     4801 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     4861 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     4921 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     4981 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     5041 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     5101 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     5161 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     5221 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     5281 ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat
     5341 agggagaccc aagcttggta ccgagctcgg atccatgaag ctgctgccgt cggtggtgct
     5401 gaagctcttt ctggctgcag ttctctcggc actggtgact ggcgagagcc tggagcggct
     5461 tcggagaggg ctagctgctg gaaccagcaa cccggaccct cccactgtat ccacggacca
     5521 gctgctaccc ctaggaggcg gccgggaccg gaaagtccgt gacttgcaag aggcagatct
     5581 ggaccttttg agagtcactt tatcctccaa gccacaagca ctggccacac caaacaagga
     5641 ggagcacggg aaaagaaaga agaaaggcaa ggggctaggg aagaagaggg acccatgtct
     5701 tcggaaatac aaggacttct gcatccatgg agaatgcaaa tatgtgaagg agctccgggc
     5761 tccctcctgc atctgccacc cgggttacca tggagagagg tgtcatgggc tgagcctccc
     5821 agtggaaaat cgcttatata cctatgacca cacaaccatc ctggccgtgg tggctgtggt
     5881 gctgtcatct gtctgtctgc tggtcatcgt ggggcttctc atgtttaggt accataggag
     5941 aggaggttat gatgtggaaa atgaagagaa agtgaagttg ggcatgacta attcccacgg
     6001 gcccttcgaa caaaaactca tctcagaaga ggatctgaat atgcatctag ggcggccgca
     6061 ctagaggaat tccgcccctc tc
//
